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1.  Relationships of Circulating Carotenoid Concentrations with Several Markers of Inflammation, Oxidative Stress, and Endothelial Dysfunction: The Coronary Artery Risk Development in Young Adults (CARDIA)/Young Adult Longitudinal Trends in Antioxidants (YALTA) Study 
Clinical chemistry  2007;53(3):447-455.
Background
Serum carotenoid concentrations relate inversely to cardiovascular disease incidence. To clarify the effect of carotenoids on atherosclerotic risk factors, we examined the association of circulating carotenoids with inflammation, oxidative stress, endothelial dysfunction, and smoking.
Methods
Black and white men and women in the Coronary Artery Risk Development in Young Adults study, ages 18 to 30 years at recruitment (1985–1986) from 4 US cities, were investigated over 15 years. We included 2048 to 4580 participants in analyses of the sum of serum α-carotene, β-carotene, zeaxanthin/lutein, and β-cryptoxanthin concentrations and of lycopene at year 0 and at year 7.
Results
The year 0 sum of 4 carotenoids was inversely associated (all P <0.05) with year 0 leukocyte count (slope per sum carotenoid SD, −0.17); year 7 fibrinogen (slope, −0.10); year 7 and year 15 C-reactive protein (slope, −0.12 and −0.09); and year 15 F2-isoprostanes (slope, −13.0), soluble P-selectin (slope, −0.48), and soluble intercellular adhesion molecule-1 (sICAM1; slope, −5.1). Leukocyte counts and sICAM1 and F2-isoprostane concentrations had stronger associations in smokers than in nonsmokers, and sICAM1 concentrations were higher in the highest carotenoid quartile in smokers than in the lowest carotenoid quartile in nonsmokers. Superoxide dismutase was positively associated with the sum of 4 carotenoids (slope, 0.12; P <0.01). Lycopene was inversely associated only with sICAM1. The year 7 carotenoid associations with these markers were mostly similar to those at year 0.
Conclusions
Circulating serum carotenoids were associated, some interactively with smoking, in apparently beneficial directions with markers of inflammation, oxidative stress, and endothelial dysfunction.
doi:10.1373/clinchem.2006.074930
PMCID: PMC2440581  PMID: 17234732
2.  Circulating Carotenoid Concentrations and Incident Hypertension: The Coronary Artery Risk Development in Young Adults (CARDIA) study 
Journal of hypertension  2009;27(2):237-242.
Background
Several epidemiological studies have demonstrated that carotenoid concentrations relate inversely to cardiovascular disease incidence. Thus, we examined the association of circulating carotenoids with hypertension, a major macrovascular disease risk factor.
Methods
Black and white men and women in the Coronary Artery Risk Development in Young Adults (CARDIA) Study, aged 18 to 30 years at recruitment (1985–1986) from 4 US cities, were investigated over 20 years. At years 0, 7, and 15, we determined the relationships of the sum of 4 serum carotenoids (α-carotene, β-carotene, lutein/zeaxanthin, cryptoxanthin) and of lycopene with incident hypertension using proportional hazards regression models.
Results
In 4412 participants, year 0 sum of 4 carotenoids was significantly inversely associated with 20 year hypertension incidence after adjustment for baseline systolic blood pressure and other confounding factors (relative hazard per SD increase of sum of 4 carotenoids: 0.91; 95% confidence interval: 0.84–0.99). The inverse relationship persisted in time-dependent models updating year 0 sum of 4 carotenoids with year 7 and year 15 values (relative hazard per SD increase of sum of 4 carotenoids: 0.84; 95% confidence interval: 0.77–0.92). Lycopene was unrelated to hypertension in any model.
Conclusion
Those with higher concentrations of sum of carotenoids, not including lycopene, generally had lower risk for future hypertension.
doi:10.1097/HJH.0b013e32832258c9
PMCID: PMC2920800  PMID: 19155781
Antioxidant; Carotenoids; Epidemiology; Hypertension; Prospective studies
3.  Carotenoid Intake and Risk of Colorectal Adenomas in a Cohort of Male Health Professionals 
Cancer causes & control : CCC  2013;24(4):705-717.
Purpose
Carotenoids have been hypothesized to prevent carcinogenesis through their antioxidant and pro-vitamin A properties. We examined associations between intakes of specific carotenoids and risk of colorectal adenomas.
Methods
Among 29,363 men who reported having a lower bowel endoscopy between 1986–2006, 3,997 cases of colorectal adenoma were identified in the Health Professionals Follow-up Study. Participants completed food frequency questionnaires every 4 years; dietary information was cumulatively updated. The associations between carotenoid intakes and risk of colorectal adenomas overall and by anatomic site, stage, smoking status and alcohol consumption were investigated using multivariate logistic regression models.
Results
Total β-carotene and dietary β-carotene, lycopene and lutein/zeaxanthin intakes and the total carotenoid score were inversely associated with colorectal adenoma risk. The odds ratios (95% confidence intervals) comparing the highest vs. lowest quintile of intake were 0.78 (0.69–0.88) for total β-carotene, 0.72 (0.64–0.81) for dietary β-carotene, 0.83 (0.74–0.93) for lycopene, 0.86 (0.76–0.96) for lutein/zeaxanthin, and 0.87 (0.77–0.97) for the total carotenoid score. Associations for α-carotene and β-cryptoxanthin intakes were null. We did not find significant differences in the associations between intakes of each carotenoid and risk of colorectal adenoma by anatomic site or stage (all p-values, test for common effects > 0.10). The inverse associations we observed for total β-carotene and dietary β-carotene, lycopene, and lutein/zeaxanthin intakes and the total carotenoid score with adenoma risk also did not vary by smoking status and alcohol consumption.
Conclusion
This study found that a diet high in carotenoids was associated with a reduced risk of colorectal adenomas.
doi:10.1007/s10552-013-0151-y
PMCID: PMC3767390  PMID: 23371557
carotenoids; nutrition; colorectal adenomas; colorectal cancer; epidemiology
4.  Associations of plasma carotenoids with risk factors and biomarkers related to cardiovascular disease in middle-aged and older women 
Background
Cardiovascular disease (CVD) risk factors may potentially influence plasma levels of carotenoids. However, data on the association of plasma carotenoids with CVD related biomarkers are only limited.
Objective
We examined the cross-sectional association of plasma carotenoids with blood lipids, hemoglobin A1c (HbA1c), and C-reactive protein (CRP) in middle-aged and older women initially free of CVD and cancer.
Design
Participants from three nested case-control studies in Women's Health Study were pooled. Baseline plasma carotenoids, including α-carotene, β-carotene, β-cryptoxanthin, lycopene, and lutein/zeaxanthin, blood lipids, HbA1c, and CRP were available for 2895 women.
Results
Women who were current smokers or obese had lower plasma levels of most carotenoids expect for lycopene. After adjusting for age, race, lifestyle factors, clinical factors, plasma total cholesterol, and dietary carotenoids, an increase of 30 mg/dl in LDL-cholesterol was associated with 17% increase in α-carotene, 16% increase in β-carotene, and 8.5% increase in lycopene; an increase of 10 mg/dl in HDL-cholesterol was associated with 5.3% decrease in lycopene; an increase of 0.3% in HbA1c was associated with 1.4% increase in lycopene; and an increase of 2 mg/L in CRP was associated with 1.3% decrease in β-carotene (all p<0.01).
Conclusions
In middle-aged and older women free of CVD and cancer, plasma carotenoids were associated with smoking, obesity, LDL-cholesterol, HDL-cholesterol, HbA1c, and CRP. The associations differ among individual carotenoids, possibly reflect metabolic effects of lifestyle and physiologic factors on plasma carotenoids, and may partially explain the inverse association of plasma carotenoids with CVD outcomes observed in epidemiologic studies.
PMCID: PMC2559966  PMID: 18779292
carotenoids; blood lipids; glycosylated hemoglobin A1c; C-reactive protein; cross-sectional study; women
5.  Low serum carotenoid concentrations and carotenoid interactions predict mortality in US adults: The Third National Health and Nutrition Examination Survey (NHANES III) 
Evidence regarding the health benefits of carotenoids is controversial. Effects of serum carotenoids and their interactions on mortality have not been examined in a representative sample of US adults. The objective was to examine whether serum carotenoid concentrations predict mortality among US adults. The study consisted of adults aged ≥20 years enrolled in the National Health and Nutrition Examination Survey (NHANES) III, 1988–1994, with measured serum carotenoids and mortality follow-up through 2006 (N=13,293). Outcomes were all-cause, cardiovascular disease (CVD), and cancer mortality. In adjusted Cox proportional hazards models, participants in the lowest total carotenoid quartile (<1.01µmol/L) had significantly higher all-cause mortality (mortality rate ratio=1.38; 95% confidence interval:1.15—1.65; P=0.005) than those in the highest total carotenoid quartile (>1.75µmol/L). For alpha-carotene, the highest quartile (>0.11µmol/L) had the lowest all-cause mortality rates (P<0.001). For lycopene, the middle two quartiles (0.29–0.58µmol/L) had the lowest all-cause mortality rates (P=0.047). Analyses with continuous carotenoids confirmed associations of serum total carotenoids, alpha-carotene, and lycopene with all-cause mortality (P<0.001). In a random survival forest analysis, very low lycopene was the carotenoid most strongly predictive of all-cause mortality, followed by very low total carotenoids. Alpha-carotene/beta-cryptoxanthin, alpha-carotene/lutein+zeaxanthin and lycopene/lutein+zeaxanthin interactions were significantly related to all-cause mortality (P<0.05). Low alpha-carotene was the only carotenoid associated with CVD mortality (P=0.002). No carotenoids were significantly associated with cancer mortality. Very low serum total carotenoid, alpha-carotene, and lycopene concentrations may be risk factors for mortality, but carotenoids show interaction effects on mortality. Interventions of balanced carotenoid combinations are needed for confirmation.
doi:10.1016/j.nutres.2011.03.003
PMCID: PMC3081783  PMID: 21481711
alpha-carotene; beta-carotene; beta-cryptoxanthin; carotenoids; human subjects; lycopene; lutein; mortality; NHANES; zeaxanthin
6.  Circulating Carotenoids and Risk of Breast Cancer: Pooled Analysis of Eight Prospective Studies 
Background
Carotenoids, micronutrients in fruits and vegetables, may reduce breast cancer risk. Most, but not all, past studies of circulating carotenoids and breast cancer have found an inverse association with at least one carotenoid, although the specific carotenoid has varied across studies.
Methods
We conducted a pooled analysis of eight cohort studies comprising more than 80% of the world’s published prospective data on plasma or serum carotenoids and breast cancer, including 3055 case subjects and 3956 matched control subjects. To account for laboratory differences and examine population differences across studies, we recalibrated participant carotenoid levels to a common standard by reassaying 20 plasma or serum samples from each cohort together at the same laboratory. Using conditional logistic regression, adjusting for several breast cancer risk factors, we calculated relative risks (RRs) and 95% confidence intervals (CIs) using quintiles defined among the control subjects from all studies. All P values are two-sided.
Results
Statistically significant inverse associations with breast cancer were observed for α-carotene (top vs bottom quintile RR = 0.87, 95% CI = 0.71 to 1.05, Ptrend = .04), β-carotene (RR = 0.83, 95% CI = 0.70 to 0.98, Ptrend = .02), lutein+zeaxanthin (RR = 0.84, 95% CI = 0.70 to 1.01, Ptrend = .05), lycopene (RR = 0.78, 95% CI = 0.62 to 0.99, Ptrend = .02), and total carotenoids (RR = 0.81, 95% CI = 0.68 to 0.96, Ptrend = .01). β-Cryptoxanthin was not statistically significantly associated with risk. Tests for heterogeneity across studies were not statistically significant. For several carotenoids, associations appeared stronger for estrogen receptor negative (ER−) than for ER+ tumors (eg, β-carotene: ER−: top vs bottom quintile RR = 0.52, 95% CI = 0.36 to 0.77, Ptrend = .001; ER+: RR = 0.83, 95% CI = 0.66 to 1.04, Ptrend = .06; Pheterogeneity = .01).
Conclusions
This comprehensive prospective analysis suggests women with higher circulating levels of α-carotene, β-carotene, lutein+zeaxanthin, lycopene, and total carotenoids may be at reduced risk of breast cancer.
doi:10.1093/jnci/djs461
PMCID: PMC3525817  PMID: 23221879
7.  Carotenoid intakes, assessed by food-frequency questionnaires (FFQs), are associated with serum carotenoid concentrations in the Jackson Heart Study: validation of the Jackson Heart Study Delta NIRI Adult FFQs 
Public health nutrition  2007;11(10):989-997.
Objectives
Intake and status of carotenoids have been associated with chronic disease. The objectives of this study were to examine the association between carotenoid intakes as measured by two regional food-frequency questionnaires (FFQs) and their corresponding measures in serum, and to report on dietary food sources of carotenoids in Jackson Heart Study (JHS) participants.
Design
Cross-sectional analysis of data for 402 African American men and women participating in the Diet and Physical Activity Sub-Study (DPASS) of the JHS.
Results
Mean serum carotenoid concentrations and intakes in this population were comparable to those reported for the general US population. After adjustment for covariates, correlations between serum and dietary measures of each carotenoid, for the average of the recalls (deattenuated), the short FFQ and the long FFQ, respectively, were: 0.37, 0.35 and 0.21 for α-carotene; 0.35, 0.26 and 0.28 for total (diet plus supplements) β-carotene; 0.25, 0.17 and 0.20 for dietary β-carotene; 0.42, 0.34 and 0.26 for β-cryptoxanthin; 0.33, 0.15 and 0.17 for lutein plus zeaxanthin; and 0.37, 0.19 and 0.14 for lycopene. Major dietary sources of α-carotene were orange vegetables; of β-carotene and lutein plus zeaxanthin, mustard, turnip and collard greens; of β-cryptoxanthin, orange juice; and of lycopene, tomato juice.
Conclusions
On average, carotenoid intakes and serum concentrations are not lower in this southern African American population than the general US population. The two regional FFQs developed for a southern US population and used as dietary assessment tools in the JHS appear to provide reasonably valid information for most of these carotenoids.
doi:10.1017/S1368980007001310
PMCID: PMC2654383  PMID: 18053294
Carotenoids; Food frequency; African American; Adult
8.  Plasma and Dietary Carotenoids Are Associated with Reduced Oxidative Stress in Women Previously Treated for Breast Cancer 
Dietary carotenoids show numerous biological activities, including antioxidant activity, induction of apoptosis, and inhibition of mammary cell proliferation. Studies examining the role of carotenoid consumption in relation to breast cancer recurrence are limited and report mixed results. We tested the hypothesis that breast cancer survivors with high dietary and plasma carotenoids would show significantly lower levels of oxidative stress than breast cancer survivors with low dietary and plasma carotenoid levels. Two hundred seven postmenopausal breast cancer survivors from the Women's Healthy Eating and Living Study volunteered for this ancillary study. Dietary data were analyzed by the Arizona Food Frequency Questionnaire and plasma carotenoids α-carotene, β-carotene, lutein plus zeaxanthin, lycopene, and β-cryptoxanthin and quantified with high-performance liquid chromatography, and immunoaffinity chromatography-monoclonal antibody–based ELISAs were used to analyze the urine samples for 8-hydroxy-2′-deoxyguanosine (8-OhdG) and 8-isoprostaglandin-F2α (8-iso-PGF2α). The correlations between dietary and plasma carotenoids were 0.34 for β-carotene, 0.46 for α-carotene, 0.39 for β-cryptoxanthin, 0.27 for lycopene, 0.30 for lutein plus zeaxanthin, and 0.30 for total carotenoids. The 8-OHdG oxidative stress biomarker was significantly reduced at the highest quartile of total plasma carotenoid concentrations (P = 0.001) and 8-iso-PGF2α was moderately reduced (P = 0.088). Dietary carotenoid levels were not significantly associated with oxidative, stress indicators, although dietary lycopene and lutein/zeaxanthin were modestly associated with 8-OHdG levels (P = 0.054 and 0.088, respectively). Key findings include a significant inverse association between total plasma carotenoid concentrations and oxidative stress as measured by urinary 8-OHdG and a moderately significant inverse association with 8-iso-PGF2α, a protective association that was not shown for dietary carotenoid intake.
doi:10.1158/1055-9965.EPI-07-0350
PMCID: PMC2575104  PMID: 17932348
9.  Serum Carotenoid Concentrations in Postmenopausal Women from the United States with and without Osteoporosis 
Antioxidant defenses may be compromised in osteoporotic women. Little is known about fruit and vegetable or carotenoid consumption among postmenopausal women. The primary carotenoids in human serum are α- and β-carotene, lycopene, β-cryptoxanthin, lutein, and zeaxanthin. This study investigated the interrelationships among serum carotenoid concentrations, fruit and vegetable intake, and osteoporosis in postmenopausal women (n = 59, 62.7 ± 8.8 y). Bone density was assessed by dual energy x-ray absorptiometry and osteoporosis diagnosis was based upon T-scores. Serum samples (n = 53) and 3-day diet records (n = 49) were analyzed. Logistic regression analyzed differences between carotenoids after adjusting for serum retinol; supplement usage; milk, yogurt, fruit, and vegetable intake; and BMI. Pearson statistics correlated carotenoids with specific fruit or vegetable intake. Serum lycopene concentrations were lower in the osteoporosis group than controls (p = 0.03). β-Cryptoxanthin intake was higher in the osteoporosis group (p = 0.0046). Total fruit and vegetable intakes were correlated with serum lycopene and β-cryptoxanthin (p = 0.03, 0.006, respectively). Serum α-carotene concentration was associated with carrot intake, and zeaxanthin and β-cryptoxanthin with lettuce intake. Carotenoids that may have beneficial skeletal effects are lower in women with osteoporosis. Research is needed to identify potential protective mechanisms or utilization of carotenoids during osteoporosis.
doi:10.1024/0300-9831.78.3.105
PMCID: PMC2692339  PMID: 19003732
β-cryptoxanthin; carotenoids; lycopene; osteoporosis; postmenopausal women
10.  Association of Serum Carotenoid Levels With Urinary Albumin Excretion in a General Japanese Population: The Yakumo Study 
Journal of Epidemiology  2013;23(6):451-456.
Background
Albuminuria is a risk factor for not only nephropathy progression but also cardiovascular disease. Oxidative stress may have a role in the positive association between albuminuria and cardiovascular disease.
Methods
This cross-sectional study investigated the associations of serum levels of carotenoids, which are dietary antioxidants, with albuminuria among 501 Japanese adults (198 men, mean age ± SD: 66.4 ± 10.0 years; 303 women, mean age ± SD: 65.4 ± 9.8 years) who attended a health examination. Serum levels of carotenoids were determined by high-performance liquid chromatography. Logistic regression analysis was used to estimate odds ratios (ORs) with 95% CIs for albuminuria after adjustment for age, body mass index, smoking habits, drinking habits, hypertension, diabetes mellitus, and dyslipidemia.
Results
Prevalence of albuminuria was 15.4% among men and 18.1% among women. Among women with albuminuria, geometric mean serum levels of canthaxanthin, lycopene, β-carotene, total carotenes, and provitamin A were significantly lower than those of normoalbuminuric women. Adjusted ORs for albuminuria among women in the highest tertiles of serum β-carotene (OR, 0.45; 95% CI, 0.20–0.98) and provitamin A (OR, 0.45; 95% CI, 0.20–0.97) were significantly lower as compared with those for women in the lowest tertile. There were no associations between serum carotenoids and albuminuria in men.
Conclusions
An increased level of serum provitamin A, especially serum β-carotene, was independently associated with lower risk of albuminuria among Japanese women.
doi:10.2188/jea.JE20130058
PMCID: PMC3834283  PMID: 24097248
carotenoids; albuminuria; cross-sectional study
11.  Serum Retinol and Carotenoids in Association with Biomarkers of Insulin Resistance among Premenopausal Women 
ISRN nutrition  2012;2013:619516-.
Objective
The aim of this study was to investigate how serum retinol and carotenoids (β-carotene, β-cryptoxanthin, lutein/zeaxanthin, lycopene) are associated with biomarkers of insulin resistance.
Research Methods and Procedures
The BioCycle Study (2005–2007) is a prospective cohort of 259 healthy premenopausal women. Fasting serum samples were collected at up to sixteen clinic visits, from which retinol, carotenoids, insulin, glucose, and sex hormone-binding globulin (SHBG) were measured. Insulin resistance was estimated by the homeostasis model assessment (HOMA-IR). Linear mixed models were used to determine associations adjusting for age, race, body mass index (BMI), education, smoking, physical activity, triglycerides, and energy intake.
Results
Retinol was positively associated with HOMA-IR (β = 0.19 (95% CI: 0.07, 0.32)) units per ug/mL increase in retinol; the relationship was driven by insulin (β = 0.20 (95% CI: 0.08, 0.31)). Retinol was inversely associated with SHBG (β = −0.22 (95% CI: −0.28, −0.16)). Although no significant associations were found between serum carotenoids and HOMA-IR, β-carotene was positively associated with SHBG and β-cryptoxanthin inversely with fasting plasma glucose.
Conclusion
Results indicate a possible role for serum retinol in the pathogenesis of type 2 diabetes. However, they do not support a strong association between individual or total serum carotenoids and insulin resistance.
doi:10.5402/2013/619516
PMCID: PMC3737585  PMID: 23936766
12.  Serum Retinol and Carotenoids in Association with Biomarkers of Insulin Resistance among Premenopausal Women 
ISRN Nutrition  2012;2013:619516.
Objective. The aim of this study was to investigate how serum retinol and carotenoids (β-carotene, β-cryptoxanthin, lutein/zeaxanthin, lycopene) are associated with biomarkers of insulin resistance. Research Methods and Procedures. The BioCycle Study (2005–2007) is a prospective cohort of 259 healthy premenopausal women. Fasting serum samples were collected at up to sixteen clinic visits, from which retinol, carotenoids, insulin, glucose, and sex hormone-binding globulin (SHBG) were measured. Insulin resistance was estimated by the homeostasis model assessment (HOMA-IR). Linear mixed models were used to determine associations adjusting for age, race, body mass index (BMI), education, smoking, physical activity, triglycerides, and energy intake. Results. Retinol was positively associated with HOMA-IR (β = 0.19 (95% CI: 0.07, 0.32)) units per ug/mL increase in retinol; the relationship was driven by insulin (β = 0.20 (95% CI: 0.08, 0.31)). Retinol was inversely associated with SHBG (β = −0.22 (95% CI: −0.28, −0.16)). Although no significant associations were found between serum carotenoids and HOMA-IR, β-carotene was positively associated with SHBG and β-cryptoxanthin inversely with fasting plasma glucose. Conclusion. Results indicate a possible role for serum retinol in the pathogenesis of type 2 diabetes. However, they do not support a strong association between individual or total serum carotenoids and insulin resistance.
doi:10.5402/2013/619516
PMCID: PMC3737585  PMID: 23936766
13.  Plasma carotenoids are associated with socioeconomic status in an urban Indigenous population: an observational study 
BMC Public Health  2011;11:76.
Background
Indigenous Australians experience poorer health than other Australians. Poor diet may contribute to this, and be related to their generally lower socioeconomic status (SES). Even within Indigenous populations, SES may be important. Our aim was to identify factors associated with plasma carotenoids as a marker of fruit and vegetable intake among urban dwelling Indigenous Australians, with a particular focus on SES.
Methods
Cross sectional study in urban dwelling Indigenous Australians participating in the DRUID (Darwin Region Urban Indigenous Diabetes) Study. An SES score, based on education, employment, household size, home ownership and income was computed and plasma carotenoids measured by high performance liquid chromatography in 897 men and women aged 15 - 81 years (mean 36, standard deviation 15). Linear regression analysis was used to determine the relationship between SES and plasma carotenoids, adjusting for demographic, health and lifestyle variables, including frequency of intakes of food groups (fruit, vegetables, takeaway foods, snacks and fruit/vegetable juice).
Results
SES was positively associated with plasma concentrations of lutein/zeaxanthin (p trend <0.001), lycopene (p trend = 0.001), α- and ß-carotene (p trend = 0.019 and 0.026 respectively), after adjusting for age, sex, glucose tolerance status, smoking, alcohol use, hypercholesterolemia, dyslipidemia, self-reported health, waist to hip ratio and body mass index. These associations remained after adjustment for self-reported frequency of intake of fruit, vegetables, takeaway foods and fruit juice, which all showed some association with plasma carotenoids. Even in the highest SES quintile, concentrations of all carotenoids (except lycopene) were lower than the mean concentrations in a non-Indigenous population.
Conclusions
Even within urban Indigenous Australians, higher SES was associated with higher concentrations of plasma carotenoids. Low plasma carotenoids have been linked with poor health outcomes; increasing accessibility of fruit and vegetables, as well as reducing smoking rates could increase concentrations and otherwise improve health, but our results suggest there may be additional factors contributing to lower carotenoid concentrations in Indigenous Australians.
doi:10.1186/1471-2458-11-76
PMCID: PMC3040145  PMID: 21288365
14.  The effect of serum carotenoids on atrophic gastritis among the inhabitants of a rural area in Hokkaido, Japan 
A total of 206 residents (76 males and 130 females) of a rural area of Hokkaido, Japan, attending a health check in August, 1997, were studied to assess the relationship between serum carotenoids and atrophic gastritis (AG). Of the participants, 91 had AG as indicated by their serum levels of pepsinogen I and pepsinogen II. Logistic regression analysis, after adjusting for gender and age, revealed that the odds ratios for serum carotenoid levels were lower for subjects with high serum levels of α-carotene (odds ratio, 0.41; 95% C.I., 0.19–0.88) and β-carotene (odds ratio, 0.41; 95% C.I., 0.18–0.91) than for those with low serum carotenoid levels. In addition, the odds ratios of subjects with high serum levels of β-cryptoxanthin (odds ratio, 0.60; 95% C.I., 0.28–1.31), provitamin A (odds ratio, 0.38; 95% C.I., 0.17–0.85), and retinol (odds ratio, 0.67; 95% C.I., 0.31–1.48) were found to be lower than the odds ratios for those with low serum levels. Odds ratios for subjects with high serum zeaxanthin/lutein levels were higher than odds ratios for those with low serum levels. These results suggest that frequent intake of foods ric, in carotenoids with provitamin A activity may reduce the risk of AG.
doi:10.1007/BF02897968
PMCID: PMC2723307  PMID: 21432259
helicobacter pylori; atrophic gastritis; green-yellow vegetables; β-carotene; provitamin A
15.  Resonance Raman detection of carotenoid antioxidants in living human tissue 
Journal of biomedical optics  2005;10(6):064028.
Increasing evidence points to the beneficial effects of carotenoid antioxidants in the human body. Several studies, for example, support the protective role of lutein and zeaxanthin in the prevention of age-related eye diseases. If present in high concentrations in the macular region of the retina, lutein and zeaxanthin provide pigmentation in this most light sensitive retinal spot, and as a result of light filtering and/or antioxidant action, delay the onset of macular degeneration with increasing age. Other carotenoids, such as lycopene and beta-carotene, play an important role as well in the protection of skin from UV and short-wavelength visible radiation. Lutein and lycopene may also have protective function for cardiovascular health, and lycopene may play a role in the prevention of prostate cancer. Motivated by the growing importance of carotenoids in health and disease, and recognizing the lack of any accepted noninvasive technology for the detection of carotenoids in living human tissue, we explore resonance Raman spectroscopy as a novel approach for noninvasive, laser optical carotenoid detection. We review the main results achieved recently with the Raman detection approach. Initially we applied the method to the detection of macular carotenoid pigments, and more recently to the detection of carotenoids in human skin and mucosal tissues. Using skin carotenoid Raman instruments, we measure the carotenoid response from the stratum corneum layer of the palm of the hand for a population of 1375 subjects and develope a portable skin Raman scanner for field studies. These experiments reveal that carotenoids are a good indicator of antioxidant status. They show that people with high oxidative stress, like smokers, and subjects with high sunlight exposure, in general, have reduced skin carotenoid levels, independent of their dietary carotenoid consumption. We find the Raman technique to be precise, specific, sensitive, and well suitable for clinical as well as field studies. The noninvasive laser technique may become a useful method for the correlation between tissue carotenoid levels and risk for malignancies or other degenerative diseases associated with oxidative stress.
doi:10.1117/1.2139974
PMCID: PMC3086339  PMID: 16409093
Raman spectroscopy; carotenoids; human skin; macula; antioxidants; noninvasive detection; lutein; zeaxanthin; lycopene; β-carotene
16.  Circulating carotenoids, mammographic density, and subsequent risk of breast cancer 
Cancer research  2009;69(24):9323.
Mammographic density is one of the strongest predictors of breast cancer risk. Recently, it has been suggested that reactive oxygen species may influence breast cancer risk through its influence on mammographic density. In the current study, we addressed this hypothesis and also assessed if the association between carotenoids and breast cancer risk varies by mammographic density. We conducted a nested case-control study consisting of 604 breast cancer cases and 626 controls with prospectively measured circulating carotenoid levels and mammographic density in the Nurses’ Health Study. Circulating levels of α-carotene, β-carotene, β-cryptoxanthin, lycopene, and lutein/zeaxanthin were measured. We utilized a computer-assisted thresholding method to measure percent mammographic density. We found no evidence that circulating carotenoids are inversely associated with mammographic density. However, mammographic density significantly modified the association between total circulating carotenoids and breast cancer (P-heterogeneity=0.008). Overall, circulating total carotenoids was inversely associated with breast cancer risk (p-trend=0.01). Among women in the highest tertile of mammographic density, total carotenoids was associated with a 50% reduction in breast cancer risk (OR=0.5; 95% CI 0.3-0.8). In contrast, there was no inverse association between carotenoids and breast cancer risk among women with low mammographic density. Similarly, among women in the highest tertile of mammographic density, high levels of circulating α-carotene, β-cryptoxanthin, lycopene and lutein/zeaxanthin were associated with a significant 40-50% reduction in breast cancer risk (P-trend<0.05). Our results suggest that plasma levels of carotenoids may play a role in reducing breast cancer risk, particularly among women with high mammographic density.
doi:10.1158/0008-5472.CAN-09-1018
PMCID: PMC2820729  PMID: 19934322
Circulating carotenoids; mammographic density; antioxidants; breast cancer; oxidative stress
17.  Silencing of beta-carotene hydroxylase increases total carotenoid and beta-carotene levels in potato tubers 
BMC Plant Biology  2007;7:11.
Background
Beta-carotene is the main dietary precursor of vitamin A. Potato tubers contain low levels of carotenoids, composed mainly of the xanthophylls lutein (in the beta-epsilon branch) and violaxanthin (in the beta-beta branch). None of these carotenoids have provitamin A activity. We have previously shown that tuber-specific silencing of the first step in the epsilon-beta branch, LCY-e, redirects metabolic flux towards beta-beta carotenoids, increases total carotenoids up to 2.5-fold and beta-carotene up to 14-fold.
Results
In this work, we silenced the non-heme beta-carotene hydroxylases CHY1 and CHY2 in the tuber. Real Time RT-PCR measurements confirmed the tuber-specific silencing of both genes . CHY silenced tubers showed more dramatic changes in carotenoid content than LCY-e silenced tubers, with beta-carotene increasing up to 38-fold and total carotenoids up to 4.5-fold. These changes were accompanied by a decrease in the immediate product of beta-carotene hydroxylation, zeaxanthin, but not of the downstream xanthophylls, viola- and neoxanthin. Changes in endogenous gene expression were extensive and partially overlapping with those of LCY-e silenced tubers: CrtISO, LCY-b and ZEP were induced in both cases, indicating that they may respond to the balance between individual carotenoid species.
Conclusion
Together with epsilon-cyclization of lycopene, beta-carotene hydroxylation is another regulatory step in potato tuber carotenogenesis. The data are consistent with a prevalent role of CHY2, which is highly expressed in tubers, in the control of this step. Combination of different engineering strategies holds good promise for the manipulation of tuber carotenoid content.
doi:10.1186/1471-2229-7-11
PMCID: PMC1828156  PMID: 17335571
18.  Food Predictors of Plasma Carotenoids 
Nutrients  2013;5(10):4051-4066.
Empirical prediction models that weight food frequency questionnaire (FFQ) food items by their relation to nutrient biomarker concentrations may estimate nutrient exposure better than nutrient intakes derived from food composition databases. Carotenoids may especially benefit because contributing foods vary in bioavailability and assessment validity. Our objective was to develop empirical prediction models for the major plasma carotenoids and total carotenoids and evaluate their validity compared with dietary intakes calculated from standard food composition tables. 4180 nonsmoking women in the Nurses’ Health Study (NHS) blood subcohort with previously measured plasma carotenoids were randomly divided into training (n = 2787) and testing (n = 1393) subsets. Empirical prediction models were developed in the training subset by stepwise selection from foods contributing ≥0.5% to intake of the relevant carotenoid. Spearman correlations between predicted and measured plasma concentrations were compared to Spearman correlations between dietary intake and measured plasma concentrations for each carotenoid. Three to 12 foods were selected for the α-carotene, β-carotene, β-cryptoxanthin, lutein/zeaxanthin, lycopene, and total carotenoids prediction models. In the testing subset, Spearman correlations with measured plasma concentrations for the calculated dietary intakes and predicted plasma concentrations, respectively, were 0.31 and 0.37 for α-carotene, 0.29 and 0.31 for β-carotene, 0.36 and 0.41 for β-cryptoxanthin, 0.28 and 0.31 for lutein/zeaxanthin, 0.22 and 0.23 for lycopene, and 0.22 and 0.27 for total carotenoids. Empirical prediction models may modestly improve assessment of some carotenoids, particularly α-carotene and β-cryptoxanthin.
doi:10.3390/nu5104051
PMCID: PMC3820058  PMID: 24152746
carotenoids; vitamin A; α-carotene; β-carotene; β-cryptoxanthin; lutein/zeaxanthin; lycopene; food predictors; biomarkers
19.  Natural Genetic Variation in Lycopene Epsilon Cyclase Tapped for Maize Biofortification 
Science (New York, N.Y.)  2008;319(5861):330-333.
Dietary vitamin A deficiency causes eye disease in 40 million children each year and places 140 to 250 million at risk for health disorders. Many children in sub-Saharan Africa subsist on maize-based diets. Maize displays considerable natural variation for carotenoid composition, including vitamin A precursors α-carotene, β-carotene, and β-cryptoxanthin. Through association analysis, linkage mapping, expression analysis, and mutagenesis, we show that variation at the lycopene epsilon cyclase (lcyE) locus alters flux down α-carotene versus β-carotene branches of the carotenoid pathway. Four natural lcyE polymorphisms explained 58% of the variation in these two branches and a threefold difference in provitamin A compounds. Selection of favorable lcyE alleles with inexpensive molecular markers will now enable developing-country breeders to more effectively produce maize grain with higher provitamin A levels.
doi:10.1126/science.1150255
PMCID: PMC2933658  PMID: 18202289
20.  Genetic ablation of carotene oxygenases and consumption of lycopene or tomato powder diets modulates carotenoid and lipid metabolism in mice 
Carotene-15,15'-monooxygenase (CMO-I) cleaves β-carotene to form vitamin A while carotene-9’,10’-monooxygenase (CMO-II) preferentially cleaves non-provitamin A carotenoids. Recent reports indicate that beta-carotene metabolites regulate dietary lipid uptake while lycopene regulates peroxisome-proliferated activator receptor (PPAR) expression. To determine the physiologic consequences of carotenoids and their interactions with CMO-I and CMO-II, we characterized mammalian carotenoid metabolism, metabolic perturbations and lipid metabolism in female CMO-I−/− and CMO-II−/− mice fed lycopene or tomato-containing diets for 30 days. We hypothesized that there would be significant interactions between diet and genotype on carotenoid accumulation and lipid parameters. CMO-I−/− mice had higher levels of leptin, insulin and hepatic lipidosis, but lower levels of serum cholesterol. CMO-II−/− mice had increased tissue lycopene and phytofluene accumulation, reduced IGF-1 levels and cholesterol levels, but elevated liver lipids and cholesterol compared with WT mice. The diets did not modulate these genotypic perturbations, but lycopene and tomato powder did significantly decrease serum insulin-like growth factor-I. Tomato powder also reduced hepatic PPAR expression, independent of genotype. These data point to the pleiotropic actions of CMO-I and CMO-II supporting a strong role of these proteins in regulating tissue carotenoid accumulation and the lipid metabolic phenotype, as well as tomato carotenoid-independent regulation of lipid metabolism.
doi:10.1016/j.nutres.2013.07.007
PMCID: PMC3804893  PMID: 24034573
Lycopene; phytoene; tomato; lipids; cholesterol; CMO-I; CMO-II; mice
21.  Associations of serum vitamin A and carotenoid levels with markers of prostate cancer detection among US men 
Cancer causes & control : CCC  2011;22(11):1483-1495.
Associations of serum vitamin A and carotenoid levels with markers of prostate cancer detection were evaluated among 3927 U.S. men, 40–85 years of age, who participated in the 2001–2006 National Health and Nutrition Examination Surveys. Five recommended definitions of prostate cancer detection were adopted using total and free prostate specific antigen (tPSA and fPSA) laboratory measurements. Men were identified as high-risk based on alternative cut-offs, namely, tPSA>10 ng/ml, tPSA>4 ng/ml, tPSA> 2.5 ng/ml, %fPSA<25% and %fPSA<15%. %fPSA was defined as (fPSA÷tPSA)×100%. Serum levels of vitamin A (retinol, retinyl esters) and carotenoids (α-carotene, β-carotene, β-cryptoxanthin, lutein+zeaxanthin, lycopene) were defined as quartiles and examined as risk/protective factors for PSA biomarkers. Odds ratios (OR) and 95% confidence intervals (CI) were estimated using binary logistic models. After adjustment for known demographic, socioeconomic and lifestyle confounders, high serum levels of retinyl esters (tPSA>10 ng/ml: Q4vs.Q1→OR=0.38, 95% CI: 0.14–1.00) and α-carotene (%fPSA<15%: Q4vs.Q1→OR=0.49, 95% CI: 0.32–0.76) were associated with a lower odds whereas high serum level of lycopene (tPSA>2.5 ng/ml: Q4vs.Q1→OR=1.49, 95% CI: 1.01–2.14) was associated with a greater odds of prostate cancer detection. Apart from the three significant associations observed, no other exposure-outcome association was significant. Monitoring specific antioxidant levels may be helpful in early detection of prostate cancer.
doi:10.1007/s10552-011-9822-8
PMCID: PMC3443554  PMID: 21800039
vitamin A; carotenoids; prostate cancer; prostate-specific antigen
22.  Serum carotenoids and vitamins and risk of cervical dysplasia from a case–control study in Japan 
British Journal of Cancer  1999;81(7):1234-1237.
The relationships between risk of cervical dysplasia and dietary and serum carotenoids and vitamins were investigated in a case–control study. Cases were 156 women who attended Papanicolaou test screening in nine institutes affiliated with Japan Study Group of Human Papillomavirus (HPV) and Cervical Cancer and had cervical dysplasia newly histologically confirmed. Age-matched controls were selected from women with normal cervical cytology attending the same clinic. Blood sample and cervical exfoliated cells were obtained for measuring serum retinol, α-carotene, β-carotene, zeaxanthin/lutein, cryptoxanthin, lycopene and α-tocopherol and for HPV detection. Higher serum level of α-carotene was significantly associated with decreased risk of cervical dysplasia after controlling for HPV infection and smoking status (odds ratio (OR) = 0.16, 95% confidence interval (CI) 0.04–0.62 for the highest as compared with the lowest tertile). Decreased risk for the highest tertile of serum lycopene (OR = 0.28) was marginally significant. Decreased risks observed for the highest tertiles of β-carotene (OR = 0.65) and zeaxanthin/lutein (OR = 0.53), were not statistically significant. © 1999 Cancer Research Campaign
doi:10.1038/sj.bjc.6690834
PMCID: PMC2374334  PMID: 10584887
cervical dysplasia; carotenoids; human papillomavirus; diet
23.  Passive smoking and chronic obstructive pulmonary disease: cross-sectional analysis of data from the Health Survey for England 
BMJ Open  2011;1(2):e000153.
Objectives
There is increasing evidence that passive smoking is associated with chronic respiratory diseases, but its association with chronic obstructive pulmonary disease (COPD) requires more study. In this cross-sectional analysis of data from 3 years of the Health Survey for England, the association between passive smoking exposure and risk of COPD is evaluated.
Design
Cross-sectional analysis of the 1995, 1996 and 2001 Health Surveys for England including participants of white ethnicity, aged 40+ years with valid lung function data. COPD was defined using the lower limit of normal spirometric criteria for airflow obstruction. Standardised questions elicited self-reported information on demography, smoking history, ethnicity, occupation, asthma and respiratory symptoms (dyspnoea, chronic cough, chronic phlegm, wheeze). Passive smoking was measured by self-report of hours of exposure to cigarette smoke per week.
Results
Increasing passive smoke exposure was independently associated with increased risk of COPD, with adjusted OR 1.05 (95% CI 0.93 to 1.18) for 1–19 h and OR 1.18 (95% CI 1.01 to 1.39) for 20 or more hours of exposure per week. Similar patterns (although attenuated and non-significant) were observed among never smokers. More marked dose–response relationships were observed between passive smoking exposure and respiratory symptoms, but the most marked effects were on the development of clinically significant COPD (airflow obstruction plus symptoms), where the risk among never smokers was doubled (OR 1.98 (95% CI 1.03 to 3.79)) if exposure exceeded 20 h/week.
Conclusion
This analysis adds weight to the evidence suggesting an association between passive smoking exposure and COPD.
Article summary
Article focus
Passive exposure to cigarette smoke is established as an important independent risk factor for the development of chronic conditions such as heart disease and lung cancer.
Although there is growing evidence implicating passive smoking in asthma and other respiratory diseases, the evidence for its effect on chronic obstructive pulmonary disease (COPD) is inconsistent.
Using cross-sectional data from the annual Health Survey for England, we examined the association between self-reported exposure to passive smoking and COPD.
Key messages
We have demonstrated a significant dose–response relationship between hours of exposure to passive smoking and increasing risk of COPD.
The most marked effects were observed on the development of clinically significant COPD (airflow obstruction plus symptoms), where the risk among never smokers was doubled (OR 1.98 (95% CI 1.03 to 3.79)) if exposure exceeded 20 h/week.
Passive smoking is prevalent worldwide, and even after the 2007 public smoking ban in the UK, 20% of the adult English population are still exposed to up to 20 h of passive smoking per week, with 5% exposed to more than 20 h/week; further measures are needed to investigate and reduce exposures in the home and elsewhere.
Strengths and limitations of this study
Our study has the advantage of being a large sample representative of the English population (>21 000 participants), conducted over 3 separate years, with a standardised protocol and objective measure of lung function.
However, due to the cross-sectional nature of the design, temporal associations cannot necessarily be inferred.
The Health Survey for England was not designed for the specific analyses presented in this paper, and thus some of the measures are crude.
Self-reported passive smoke exposure is only a proxy for true exposure levels, but is accepted as the most practical method of assessment.
doi:10.1136/bmjopen-2011-000153
PMCID: PMC3191589  PMID: 22021874
24.  A Mediterranean Dietary Intervention in Healthy American Women Changes Plasma Carotenoids and Fatty Acids in Distinct Clusters 
This study examined patterns of changes in plasma fatty acids and carotenoids when women were asked to follow a novel, Greek-Mediterranean exchange list diet. A total of 69 healthy, non-obese, women ages 25–59, were randomized to either continue their own usual diet or to follow a modified Mediterranean diet for six months. There were no significant changes in blood lipids, triacylglycerol, insulin, glucose or C-reactive protein. Mean plasma carotenoids increased by 55%, which is consistent with a large increase in fruit and vegetable consumption. Likewise, changes in fat intakes were reflected in blood fatty acids, with a 25% increase in mean plasma monounsaturated fatty acids (MUFA). Principal component analysis was conducted to examine the sources of inter-individual variation for changes in carotenoid and fatty acid levels. Changes in the Mediterranean diet were clustered together in four components that accounted for 78% of the variance in plasma levels. Increases in plasma lutein, α- and β-carotene clustered together in a “vegetable” pattern, and increases in carotenoids found in fruit, β-cryptoxanthin and zeaxanthin, also clustered together but accounted for less of the variance. Increases in plasma MUFA were clustered with a decrease in plasma polyunsaturated fatty acids, consistent with substitution in the type of oils consumed. The only association of fatty acid levels with carotenoids was that of lycopene, which clustered together with an increase in saturated fatty acids. The changes in blood levels indicate the exchange list diet was effective for targeting Mediterranean nutrient intakes using foods available in the United States.
doi:10.1016/j.nutres.2009.03.001
PMCID: PMC2735788  PMID: 19358929
cancer prevention; monounsaturated fat; fruit; vegetables; olive oil; human; clinical trial; carotenoids
25.  Changes in the concentrations of biochemical indicators of diet and nutritional status of pregnant women across pregnancy trimesters in Trujillo, Peru, 2004–2005 
Nutrition Journal  2013;12:80.
Background
In developing countries, deficiencies in essential micronutrients are common, particularly in pregnant women. Although, biochemical indicators of diet and nutrition are useful to assess nutritional status, few studies have examined such indicators throughout pregnancy in women in developing countries.
Methods
The primary objective of this study was to assess the nutritional status of 78 Peruvian women throughout pregnancy for 16 different nutritional indicators including fat-soluble vitamins and carotenoids, iron-status indicators, and selenium. Venous blood samples from which serum was prepared were collected during trimesters one (n = 78), two (n = 65), three (n = 62), and at term via the umbilical cord (n = 52). Questionnaires were completed to determine the demographic characteristics of subjects. Linear mixed effects models were used to study the associations between each maternal indicator and the demographic characteristics.
Results
None of the women were vitamin A and E deficient at any stage of pregnancy and only 1/62 women (1.6%) was selenium deficient during the third trimester. However, 6.4%, 44% and 64% of women had ferritin levels indicative of iron deficiency during the first, second and third trimester, respectively. Statistically significant changes (p ≤ 0.05) throughout pregnancy were noted for 15/16 nutritional indicators for this Peruvian cohort, with little-to-no association with demographic characteristics. Three carotenoids (beta-carotene, beta-cryptoxanthin and trans-lycopene) were significantly associated with education status, while trans-lycopene was associated with age and beta-cryptoxanthin with SES (p < 0.05). Concentrations of retinol, tocopherol, beta-cryptoxanthin, lutein + zeaxanthin and selenium were lower in cord serum compared with maternal serum (p < 0.05). Conversely, levels of iron status indicators (ferritin, transferrin saturation and iron) were higher in cord serum (p < 0.05).
Conclusion
The increasing prevalence of iron deficiency throughout pregnancy in these Peruvian women was expected. It was surprising though not to find deficiencies in other nutrients. The results highlight the importance of continual monitoring of women throughout pregnancy for iron deficiency which could be caused by increasing fetal needs and/or inadequate iron intake as pregnancy progresses.
doi:10.1186/1475-2891-12-80
PMCID: PMC3685542  PMID: 23758715
Micronutrients; Pregnant women; Trimester; Serum; Cord blood; Peru

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